陆地棉基因ghwrky40-1的克隆及表达分析

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1、棉花学报CottonScience2017,29(2):166―176陆地棉基因40-1的克隆及表达分析司爱君袁田琴袁陈红袁余渝渊新疆农垦科学院棉花研究所/农业部西北内陆区棉花生物学与遗传育种重点实验室/新疆兵团棉花遗传改良与高产栽培重点实验室,新疆石河子832000冤摘要院揖目的铱WRKY转录因子通过激活下游一系列抗逆应答基因的表达而提高植株的综合抗性遥本研究旨在研究WRKY转录因子在陆地棉中的功能遥揖方法铱通过基因芯片筛选鉴定出1个逆境诱导的陆地棉WRKY转录因子基因袁由于此基因编码蛋白含有1个典型的WRKY结构域袁且与拟南芥40具

2、有较高相似性袁故命名为40-1遥采用电子克隆及反转录-聚合酶链反应技术获得40-1基因cDNA全长袁并对其进行序列分析及结构与功能预测遥揖结果铱40-1的开放阅读框为981bp袁编码326个氨基酸袁预测蛋白相对分子质量为35.78伊103袁等电点为8.39遥GhWRKY40-1在拟南芥原生质体中瞬时表达定位于细胞核袁且具有转录激活活性遥RT-PCR结果表明袁40-1受甘露醇诱导上调表达遥揖结论铱推测40-1可能参与干旱胁迫应答反应袁并且在植物逆境胁迫中发挥重要作用遥上述分析为进一步从分子水平上验证其抗逆生物学功能以及揭示棉花非生物胁迫抗

3、逆机制奠定了基础遥关键词院棉花曰WRKY曰转录因子曰基因克隆曰序列分析中图分类号院S562.035文献标志码院A文章编号院1002-7807渊2017冤02-0166-1110.11963/issn.1002-7807.201702006SiAijun,TianQin,ChenHong,YuYu(832000,)[Objective]WRKYtranscriptionfactorsenhanceplantresistancebyactivatingtheexpressionofaseriesofstress-re-sponsivedow

4、nstreamgenes.ThepurposeofourresearchwastoanalyzefunctionsofWRKYtranscriptionfactorsinuplandcot-ton.[Method]Astress-inducedupland-cottonWRKYtranscriptionfactorgenewasscreenedbycDNAmicroarrayanalysis.Thegene,whichwasfoundtocontainatypicalWRKYdomainandtoshareahighlysimilara

5、minoacidsequencewith40,wasaccordinglynamed40-1.Thefull-lengthcDNAofGhWRKY40-1wasobtainedbyelectroniccloningandreversetranscription-polymerasechainreaction(RT-PCR)technology,withproteinsequenceanalysisandstructure-functionpredictionperformedusingbioinformaticssoftware.[Re

6、sult]Accordingtoourresults,40-1containsa981-bpopenreadingframeencoding326aminoacidresidues,withapredictedrelativemolecularmassof35.78kDaandanisoelectricpointof8.39.Inprotoplasts,GhWRKY40-1waslocalizedinthenucleusandwasfoundtohavetranscriptionalactivationactivity.RT-PCRan

7、alysisshowedthat40-1wasup-regulatedundermannitolstress.[Conclusion]Wehypothesizethat40-1isinvolvedindroughtstressresponseandplaysanimportantroleinplantstressresponses.Ourcharacterizationandbioinformaticsanalyseshavelaidthefoundationforfutureconfirmationofthebiologicalfun

8、ctionsofresistanceatthemolecularlevelandelucidationofmechanismsofabioticstressresistanceincotton.cotton